Antimicrobial composition

ABSTRACT

An antiseptic composition suitable for use on skin and wounds comprising a source of an antimicrobial agent and an agent which disrupts biofilms. More, particularly the invention relates to a composition capable of providing effective antimicrobial activity while at the same time avoiding wound and skin irritation and retardation of wound healing.

This invention relates to an antimicrobial composition which can beapplied to skin, wounds, cuts, abrasions or burns for the prevention ortreatment of infections. More particularly, the invention relates to acomposition capable of providing effective antimicrobial activity whileat the same time avoiding wound and skin irritation and retardation ofwound healing.

BACKGROUND OF THE INVENTION

Overuse of antibiotics and the associated increase in bacterialresistance is impacting the efficacy of antibiotics in the treatment ofwound infection. Effective alternatives to antibiotics are thusdesirable.

Topical antimicrobial materials, and preparations containing them, havelong been recognized as playing an important part in minimizing theopportunity for skin and wound infections. Antiseptics are non-selectivechemical agents that can be safe to use on living tissue. Moleculariodine, ionic silver and oxidizing agents, such as sodium hypochloriteand chlorine dioxide, have been recognized as antiseptic agents witheffectiveness against a wide range of microorganisms. There are,however, several barriers to making an effective antimicrobialcomposition for application to wounds based on such agents. One problemis that these antiseptic agents tend to react with organic materialsfound in the wound other than the intended microbial targets. This meansthat to be effective, antiseptic agents need to be included in treatmentcompositions at high levels, which may cause undesirable side effectswith prolonged use such as cell toxicity, hypersensitivity reactions,skin staining and systemic effects. Such side effects are furtherdescribed in “In vitro cytotoxity of silver: implication for clinicalwound care”. Poon V K, Burd A. Burns. March 2004; 30(2):140-7, “A reviewof iodine toxicity reports”. Pennington J A. J Am Diet Assoc. November1990; 90(11):1571-81 and “Topical antimicrobial toxicity”. LineaweaverW, Howard R, Soucy D, McMorris S, Freeman J, Crain C, Robertson J,Rumley T. Arch Surg. March 1985; 120(3):267-70.

Wounds are often colonized by a variety of microorganisms, some of whichmay cause infection. It is increasingly recognized that microbialpopulations living within a biofilm environment contribute to delayedhealing and infection. Biofilms are comprised of exopolymeric substancesthat are produced by bacteria once the bacteria attach to a surface, andthis helps to protect microorganisms from immune cells and antimicrobialagents. Since efficacy of antimicrobial agents (e.g., antibiotics andantiseptics) is compromised by the biofilm matrix, strategies to disruptthe biofilm and expose microorganisms within can be helpful inincreasing the activity level of antimicrobial agents and thus reducethe concentration of such agents needed to make an effectivecomposition.

Ethylenediaminetetra-acetic acid (EDTA) added as the di-sodium orcalcium di-sodium salts has been used to treat topical infections or totreat hard surfaces such as catheters. WO03/047341 describes the use ofEDTA, for example, as an additive for a toothpaste. EDTA is also used asa formulation agent to reduce the effects of water hardness and,generally, as a chelating agent.

EDTA has been described in combination with other antibiotic agents. Forexample, in U.S. Pat. No. 5,998,488, EDTA is used in combination with anantimicrobial preservative in a solution for ophthalmic use.Formulations suitable for topical use on wounds have not been proposed.

There is, thus, a need for a composition which has the benefits of anantimicrobial agent but which reduces the potential of adversereactions. Surprisingly, we have found that EDTA is capable ofdisrupting biofilms by chelating the metal ions, calcium and magnesium,that maintain the integrity of the biofilm matrix.

SUMMARY OF THE INVENTION

Accordingly, a first aspect of the invention provides an antimicrobialcomposition suitable for use on skin and wounds comprising a source ofantiseptic agent and an agent which disrupts biofilms such as EDTA.

The presence of EDTA enhances the effect of the antiseptic so that theconcentration of antiseptic agent may be reduced and yet still achieveeffective antisepsis. By increasing the effectiveness of the antisepticagent, its concentration in the composition can be reduced, thereby,reducing the potential for adverse reactions.

This aspect of the invention allows the preparation of compositions ableto reduce the bacterial bioburden to a level manageable by the hostwithin 24 hours using at least 50% less of the antiseptic agent,preferably 50-60% less, and, more preferably, 65-85% less. This thenallows the wound to progress towards healing and can been evidenced by avisual improvement in the wound.

We have also found that it is possible to prepare a composition whichincludes EDTA which is effective under the conditions of pH normallyfound in a wound.

Accordingly, a second aspect of the invention provides a compositionsuitable for use on wounds comprising di-, tri- and tetra-basic salts ofEDTA at a pH of 4 to 8.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the antimicrobial activity of a two componentiodine generating solution with and without EDTA against Staphylococcusaureus.

FIG. 2 is a graph showing zone of inhibition with EDTA.

FIG. 3 is a graph showing zones of inhibition with EDTA, with andwithout poloaxmer.

DETAILED DESCRIPTION OF THE INVENTION

The compositions according to a first aspect of the invention comprisean antiseptic agent, preferably molecular iodine, ionic silver,chlorhexidine, or hydrochloric acid or an oxidizing agent such as sodiumhypochlorite, chlorine dioxide, hydrogen peroxide or peroxy acid salts.The antiseptic agent is preferably included in the composition at alevel of from 0.01% to 10% by weight, more preferably 0.1% to 5% byweight. Iodine is preferably included in the composition at a level offrom 0.01% to 10% by weight and more preferably from 0.1% to 1.0% byweight. Preferably, the source of iodine is an iodide and thecomposition further comprises an oxidant and a buffer, the oxidant beingheld separately from the iodide until the point of use. The buffer ispreferably capable of maintaining the pH of the composition at between4.5 and 6 so that iodine is generated at a physiologically acceptableand efficacious rate. Compositions comprising iodide and an oxidant heldseparately from the iodide are described further in EP1158859B.

Where iodide is present, the amount of oxidant in the composition istailored to provide a stoichiometric match with iodide. Preferably, theoxidant is iodate and is provided in a molar ratio of from 1:4 to 1:10with iodide. In this way, the iodide present in the composition fullyreacts with the oxidant. Iodide and iodate are preferably present assodium salts although other usual counter ions may be present.

Where the antiseptic agent is ionic silver, it is preferably included inthe composition at a level of from 0.1% to 10% by weight and morepreferably 0.5% to 1.5% by weight.

The pH of the composition is generally below 8 and preferably between 4and 8, more preferably between 4 and 6 and most preferably between 4.5and 5.5. The desired pH may be achieved by incorporating bufferingagents in the composition. Examples of buffering agents which may beincluded are citric acid/di-sodium hydrogen phosphate, citricacid/sodium citrate, acetic acid/sodium acetate. The buffering agent mayconveniently be present in an amount of about 1% to 20% by weight of thecomposition, preferably about 4% to 6% by weight and more preferablyabout 5% by weight so as to provide an isotonic composition

EDTA is preferably present as the di-, tri- or tetra-basic salts ofEDTA. We have found that these salts are effective for eradicatingmicroorganisms in the free floating or planktonic state and biofilmstate alone or in the presence of an antiseptic agent. For example, wehave found that EDTA at concentrations of 0.1-40% weight by volume waseffective in killing a range of microorganisms both in the planktonicand biofilm state. Microorganisms that were effectively killed by EDTAincluded Pseudomonas aeruginosa, Serratia marcescens, vancomycinresistant Enterococcus (VRE) and methicillin resistant Staphylococcusaureus (MRSA).

EDTA is preferably present in the compositions of either aspect of thepresent invention at a level of 0.5% to 10% by weight of thecomposition, more preferably 1% to 3% by weight.

The compositions of the present invention may be in the form of a waterbased gel which maintains a moist wound healing environment and promoteshealing. A gel gives the advantage of flow into the wound to form anintimate contact with the wound bed and provide antimicrobial effects tothe whole wound. Preferably, the gel has a high enough viscosity that itdoes not flow out of wounds on areas of the body that are or becomenon-horizontal. Preferably, the pH of the gel is buffered at around 5.5as this does not alter the pH balance of the peri-wound tissue and,therefore, protects it.

The following examples are illustrative of the present invention.

EXAMPLE 1

Effect of EDTA on Iodine

Compositions containing iodine and EDTA were prepared by making a pairof aqueous gels (see Table 1 for Gel A and Table 2 for Gel B) which wereintimately mixed at the point of use. Each gel was made by preparing anaqueous solution containing all of the appropriate water-soluble partsaccording to the formulations below and then adding a slurry of anon-ionic cellulosic viscosifier (hydroxyethylcellulose) in propyleneglycol. TABLE 1 Component % w/w Water To 100 Propan-1,2-diol 10Hydroxyethylcellulose 3.86 Sodium iodate 0.16 Citric acid 3.99 di-sodiumphosphate 15.06

TABLE 2 Component % w/w Water To 100 Propan-1,2-diol 10Hydroxyethylcellulose 4.14 Sodium iodide 0.59 Ethylenediaminetetraacetic0.1-4.0 acid tetra-sodium salt tetrahydrate Phosphoric acid As necessaryto adjust aqueous phase to ph 5.5

A commercially available cadexomer iodine ointment was present as apositive control as it contains 0.9% iodine. Intrasite is an amorphoushydrogel and was present as a negative control as it contains 0% iodine.

9 ml of simulated wound fluid was added to a 17 ml volume cell well. A 1ml culture of Staphylococcus aureus was added to each well to give afinal culture concentration of 10⁶ cfu/ml. Controls for this experimentinvolved adding 2 g of cadexomor iodine ointment (positive control) andnydrogel (negative control) separately to three cell wells each. 1 g ofgels A and B, with different concentrations of EDTA, were then added toseparate cell wells (in triplicate). The cell wells containing theculture and gels were then shaken at 600 rpm at 35° C. After timeintervals of 4, 24, 48, 72 and 96 hours, a 0.1 ml test sample was takenfrom each well and placed into 9.9 ml MRD (maximum recovery diluent)containing 1% sodium thiosulphate. A 1 ml sample was then transferred toTryptone Soy Agar plates and incubated for 48 hours. Bacterial countswere then recorded. TABLE 3 Test composition 4 hours 24 hours 48 hours72 hours 96 hours Iodosorb(positive 10 10 10 10 10 control)Intrasite(negative 7600000 310000000 530000000 1000000000 14000000control)   0% I₂ 0% EDTA 7600000 310000000 530000000 1000000000 14000000 0.3% I₂ 0% EDTA 1240000 1000000 1000000 10000000 100000000  0.3% I₂ 0%EDTA 1010000 1000000 1000000 10000000 100000000  0.3% I₂ 0% EDTA 16700001000000 1000000 10000000 100000000  0.3% I₂ 1500000 1000000 10000001000000 1000000  0.5% EDTA  0.3% I₂ 92000 10 10 10 10  0.5% EDTA  0.3%I₂ 58000 10 10 10 10  0.5% EDTA  0.3% I₂ 1% EDTA 8100 10 10 10 10  0.3%I₂ 1% EDTA 130000 10 10 10 10  0.3% I₂ 1% EDTA 110000 10 10 10 10  0.3%I₂ 710 10 10 10 10 1.75% EDTA  0.3% I₂ 4400 10 10 10 10 1.75% EDTA  0.3%I₂ 31000 10 10 10 10 1.75% EDTA  0.3% I₂ 8500 10 10 10 10  2.5% EDTA 0.3% I₂ 10600 10 10 10 10  2.5% EDTA  0.3% I₂ 6200 16000 10 10 10  2.5%EDTA  0.3% I₂ 4% EDTA 93000 10 10 10 10  0.3% I₂ 4% EDTA 8500 10 10 1010  0.3% I₂ 4% EDTA 1400 10 10 10The results shown in Table 3 are also shown graphically in FIG. 1.

These results show that by adding as little as 0.5% EDTA (calculated asthe di-sodium salt) to iodine, the efficacy of iodine, at 0.3%, isenhanced when compared to the control of 0.3% iodine with no EDTA.Clearly, from the results, EDTA enhances the effects of iodine within 24hours to the same efficacy of 0.9% iodine (positive control).

EXAMPLE 2

Zones of Inhibition with Tetra Sodium EDTA

Seven antibiotic resistant microorganisms were used to evaluate theefficacy of EDTA in killing bacteria and yeasts grown on agar. For thisexperiment, filter paper discs were soaked in EDTA at concentrationsranging from 0.1-40%. EDTA was made up by dissolving it in anappropriate amount of sterile double distilled water. The filters werethen added onto Muller Hinton agar which had been inoculated with amicroorganism under study for 24 hours at 35° C. All microorganisms weretested twice.

The results in FIG. 2 show that the “zones of inhibition” generallyranged from about 20 to 43 mm (including disc 13 mm). Zones were higherwith increasing levels of EDTA indicating that EDTA alone is aneffective antiseptic agent.

EXAMPLE 3

Zones of Inhibition with EDTA

Poloxamer F127 hydrogels (Univar, Basildon, Essex, UK) are di-blockco-polymers of polyoxyethylene and polyoxypropylene that demonstratethermo-reversible gelation properties. At temperatures below 15° C.,poloxamer is liquid and fully miscible with water but changes to a firmgel at temperatures in excess of 15° C. Poloxamer encourages bacteria toexhibit a more clinically relevant biofilm phenotype. In The Use ofPoloxamer Hydrogels for the Assessment of Biofilm Susceptibility TowardsBiocide Treatments. Gilbert P., Jones, M. V., Allison, D. G., Heys, S.,Maira, T., Wood, P. Journal of Applied Microbiology (1998; 85:985-990),Gilbert, et al., determined that P. aeruginosa cells grown on poloxamerhydrogel (biofilm form) express outer membrane proteins between 78 and87 kDa, which are not evident in cells grown on standard nutrient agar(‘planktonic’). Consequently, poloxamer gel cultures mimic many of theproperties of biofilm-grown P. aeruginosa (Gilbert et al., 1998). Thisindicates that there is a phenotypic difference between P. aeruginosacells grown on poloxamer hydrogel and nutrient agar, with only poloxamergrown cells resembling biofilm cells. It was found from Wirtanen's study(Performance Evaluation of Disinfectant Formulations UsingPoloxamer-hydrogel Biofilm-Constructs. Wirtanen, G., Salo, S., Allison,D. G., Mattila-Sandholm, T., Gilbert, P. Journal of Applied Microbiology(1998; 85:965-971)) that bacteria which are grown in poloxamer havebiofilm properties and associated enhanced biocide resistance. Gilbertand colleagues suggested that bacteria grown in poloxamer hydrogelscould be exposed to biocides to provide a reproducible method fortesting the antimicrobial efficacy of biocides against biofilm bacteria(Gilbert, et al., 1998).

Seven antibiotic resistant microorganisms were used to evaluate theefficacy of EDTA in killing bacteria and yeasts grown on poloxamer gel(biofilm state). For this experiment, filter paper discs were soaked inEDTA at concentrations ranging from 0.1-40%. In this present study,poloxamer F127, a di-block copolymer of polyoxyethylene andpolyoxypropylene, was used as a medium on which bacteria could be grownas a biofilm phenotype and express the characteristics more appropriateto the ‘real world’. The filters were then added onto Muller Hinton agarwhich had been inoculated with the microorganism under study for 24hours at 35° C. All plates were done in duplicate.

The results in FIG. 3 show that the zones of inhibition were slightlysmaller in the presence of poloxamer indicating that bacteria/yeastsgrowing as biofilms are physically more resistant to EDTA when comparedto their planktonic couterparts. C kruzei and Ps aeruginosa were theexception with larger zones with poloxamer. Zones of inhibition wereevident on all organisms tested at 40% EDTA. At 10% EDTA and 5% EDTAthere were no zones with Serratia marcescens. At 0.1% EDTA no zones ofinhibition were evident with any organism studied indicating that EDTAis not effective at this level.

EXAMPLE 4

Minimum Inhibitory Concentrations (MIC's) and Minimum BactericidalConcentrations (MBC's) with Tetra, Tri and Di Basic Salts of EDTA

A microtitre plate and optical density readings and visual inspectionwere performed to obtain the MIC's for a variety of microorganisms (seeTable 4). Concentrations of EDTA included in this study were 40 mg/ml pH10.00 as tetra-Na, 40 mg/ml as tri-Na pH 6.84, 40 mg/ml as di-Na pH5.50. Into each microtitre plate, 100 μl of inoculum and EDTA was added.The plate was then incubated for 24 hours at 35° C. ±3° C. Followingincubation all microtitre plates were inspected visually for growth.

The results are presented in Table 4. TABLE 4 IN- MICRO OCULUM MICsORGANISM IDENTIFICATION cfu di Na tri Na tetra Na S multi res 5.00E+040.938 1.875 0.938 marcescens Ps NCTC 8506 (res) 6.75E+04 0.234 0.2340.234 aeruginosa Ps NCIMB 8626 7.00E+04 0.938 0.938 0.938 aeruginosa Ecoli NCIMB 8545 5.50E+04 0.469 0.469 0.469 E coli NCIMB 10544 <5.0+040.469 0.938 0.469 KI pneumoniae 033 clinical isolate <5.0+E4 0.938 0.9381.875 Ent cloacae 166 clinical isolate 5.00E+04 1.875 0.938 0.938 Prmirabilis NCTC 9559 1.50E+05 0.469 0.469 0.469 A baumannii NCIMB 92144.40E+04 0.234 0.234 0.469 S aureus NCIMB 9518 4.00E+04 0.234 0.2340.234 MRSA 1 Cardiff PHL 5.50E+04 0.117 0.469 0.234 MRSA 2 Cardiff PHL5.75E+04 0.234 0.469 0.234 MRSA 26 1.25E+04 0.234 0.234 0.234 MRSA NCTC12232 5.00E+04 0.234 0.234 0.469 MRSA NCTC 10442 6.50E+04 0.234 0.2340.234 MRSA 103731 Chester 6.00E+04 0.234 0.117 0.234 PHL Ent faecalis141 clinical isolate 2.50E+04 0.234 0.234 0.234 VRE 1 Cardiff PHL2.75E+04 0.234 0.469 0.234 VRE 2 Cardiff PHL 3.25E+04 0.234 0.234 0.234VRE NCTC 12201 6.00E+04 0.234 0.234 0.234 Strep NCTC 8198 7.00E+04 0.2340.469 0.469 pyogenes B subtilis NCTC 3610 3.00E+04 0.234 0.234 0.234 Ckrusei NCPF 3876 (res) 1.50E+04 0.938 0.938 0.938 B fragilis NCIMB 93431.90E+05 0.117 0.234 0.234 CI 362 clinical isolate 9.00E+03 0.117 <0.117<0.117 perfringens Pep NCTC 11460 1.30E+06 0.234 0.469 0.469 anaerobius

In general, all MIC's recorded were equivalent for all the salts of EDTAstudied. This suggests that the pH of the solutions is equivalent.Therefore, the activity of EDTA is not affected by the salt form addedto the microtitre plate. These results show that low concentrations ofEDTA are very effective on bacteria.

EXAMPLE 5

The Effect of EDTA on the Antimicrobial Efficacy of Silver ContainingWound Dressings

The antimicrobial dressings used in this study were Acticoat™ (Smith andNephew) and AQUACEL® Ag Hydrofiber® (ConvaTec). Acticoat is ananocrystalline silver antimicrobial barrier dressing which consists ofa rayon/polyester non-woven inner core laminated between two layers ofsilver-coated high density polyethylene mesh (HDPE). The layers are heldtogether with ultrasound welds. AQUACEL® Ag is comprised of sodiumcarboxymethylcellulose Hydrofiber® and ionic silver. The silver cationsin AQUACEL® Ag are associated with the individual highly absorbentanionic carboxymethylcellulose fibres of the Hydrofiber® dressing.AQUACEL® Hydrofiber® dressing (without silver) was also used as acontrol.

All dressings (AQUACEL® Hydrofiber® dressing [control—without silver],AQUACEL® Ag and Acticoat™ (nanocrystalline dressing) were hydrated with20 mg/ml of tetra-Na EDTA. All tests were performed against Psaeruginosa and tested on Mueller Hinton agar (MHA) and Poloxamer gel(incorporating Mueller Hinton broth (MHB). This involved inoculatingeither a MH agar plate or poloxamer gel plate with a specific isolateand then adding an appropriate hydrated (to saturation point) wounddressing (360 I (MRD) for AQUACEL® and AQUACEL® Ag and 150 I (steriledistilled water—as per manufacturers instructions) for Acticoat™). Theplates were then incubated at 35° C. ±3° C for 24 hours after which thezone of clearance (no growth) around the dressing was measured. Zones ofinhibition were measured horizontally and vertically (inclusive of thedressing sample) and a mean value was calculated from the duplicate setof results. The mean dressing size was then subtracted from the meanzone of inhibition to determine the corrected zone of inhibition (CZOI).A CZOI test allows for any inherent variability in the shape and size ofzones created by the silver dressings which may change in dimensionfollowing hydration.

Table 5 shows the size of corrected zone of inhibition (CZOI) in mm onMH agar and poloxamer gel. TABLE 5 MHA Poloxamer gel with MHB(Non-biofilm bacteria) (biofilm bacteria) Hydrating AQUACEL AQUACELFluid AQUACEL Ag Acticoat AQUACEL Ag Acticoat EDTA 0 16 10 6 6 7 MRD 018 N/A 0 3 N/A Water N/A N/A  7 N/A N/A 3

When EDTA was added to AQUACEL®, no zones of inhibition were observed onMHA (non-biofilm state). However, ZOI's were observed around AQUACEL® Agon MHA (non-biofilm bacteria) when EDTA or (maximal recovery diluent)MRD was added indicating the antimicrobial activity of silver. LargerZOI's were observed around Acticoat™ following the addition of EDTA whencompared to hydration with water. In the presence of poloxamer gel anincrease in the CZOI was observed following hydration of AQUACEL® Agwith MRD when compared to EDTA indicating an additive effect with theuse of EDTA. Overall the results showed that EDTA enhances the effectsof ionic silver on bacteria grown in the biofilm state (poloxamer gel).Overall these results suggest that by using an EDTA dressing it iseffective against both planktonic and biofilm microorganisms.

Table 6 shows the size of corrected zone of inhibition in mm on MH agarand poloxamer gel. TABLE 6 MHA Poloxamer gel with MHB(biofilm(non-biofilm bacteria) bacteria) Nano- Nano- crystalline crystallineHydrating AQUACEL silver AQUACEL silver fluid AQUACEL Ag dressingAQUACEL Ag dressing EDTA 0 15.99 9.57 6.28 5.96 6.73 MRD 0 18.04 N/A 0  3.42 N/A Water N/A N/A 6.74 N/A N/A 2.82

EXAMPLE 6

Minimum Inhibitory Concentrations (MIC) for a Two Component GelComprising Iodine and a Two Component Gel as Detailed in Example 1Containing EDTA (diNa EDTA, triNa EDTA and tetraNa EDTA)

This experiment shows the effect of pH on the activity of a range ofEDTA forms.

For this experiment a two component gel as detailed in Example 1 wasmade without the HEC component (gel). This experiment was thereforeperformed using only liquid compositions (see Table 7 for TCG Solution(A) and Table 8 for TCG Solution (B)) in order for MIC's to becalculated. The pH of the TCG/EDTA solutions was approx 5.5. Theorganisms tested included: Staphylococcus aureus, Pseudomonasaeruginosa, Escherichia coli (repeated twice) and C albicans. TABLE 7Component % w/w Water To 100 Sodium iodate 0.1 Citric acid 3.99di-sodium phosphate 15.06

TABLE 8 Component % w/w Water To 100 Sodium iodide 0.4Ethylenediaminetetraacetic As shown in Table 1 acid tetra-sodium salttetrahydrate Phosphoric acid As necessary to adjust aqueous phase to pH5.5

Table 9 shows MIC's for TCG solution and TCG solution combined with EDTA(values in brackets are EDTA concentrations in mg/ml) TABLE 9 TCG TCGand tri TCG and tetra Bacterium solution* TCG EDTA Na EDTA Na EDTA Saureus 0.25 (0) 0.008 (0.31) 0.008 (0.31) 0.008 (0.31) Ps  0.5 (0) 0.062(2.5)  0.062 (2.5)  0.062 (2.5)  aeruginosa E coli 0.25 (0) 0.031 (1.25)0.031 (1.25) 0.031 (1.25)*(working concentration was 0.2% iodine)

MIC's were reduced considerably in the presence of EDTA (di, tri andtetra Na). It can be seen that the MICs, expressed as Iodineconcentration, are lower in the presence of di, tri and tetra sodiumEDTA.

The antimicrobial benefit of having EDTA present is achieved atconcentrations of 0.31 mg/ml. Despite different forms of EDTA beingknown to provide differential antimicrobial efficacy, at a constant pH(5.5 in this case) all EDTA forms were equally effective insignificantly reducing the MIC's for all three organisms compared to theiodine generating solution without EDTA.

1. An antiseptic composition suitable for use on skin and woundscomprising a source of an antimicrobial agent and an agent whichdisrupts biofilms.
 2. The antiseptic composition as claimed in claim 1wherein the agent which disrupts biofilms is EDTA.
 3. The antisepticcomposition as claimed in claim 1 wherein the antimicrobial agent isselected from the group consisting of iodine, ionic silver or anoxidizing agent.
 4. The antiseptic composition as claimed in claim 1wherein the EDTA is in the form of the di-, tri- or tetra-basic salts ofEDTA at a pH of about 4 to
 8. 5. The antiseptic composition as claimedin claim 1 wherein the composition has a pH of between about 4 and
 8. 6.The antiseptic composition as claimed in claim 5 wherein the compositionhas a pH of between about 4.5 and 5.5.
 7. The antiseptic composition asclaimed in claim 1 wherein the composition is in the form of a gel. 8.The antiseptic composition as claimed in claim 3 wherein theantimicrobial agent is iodine.
 9. The antiseptic composition as claimedin claim 3 wherein the composition comprises an iodine source, anoxidant and a buffer wherein the iodine is held separately from theoxidant until the point of use.
 10. The antiseptic composition asclaimed in claim 9 wherein the composition is capable of generating fromabout 5 μg of iodine per gram of composition per hour to about 1500 μgof iodine per gram of composition per hour.
 11. The antisepticcomposition as claimed in claim 3 wherein the antimicrobial agent isionic silver.
 12. The antiseptic composition as claimed in claim 11wherein the composition comprises from about 0.1% to about 10% by weightof ionic silver.
 13. The antiseptic composition as claimed in claim 13wherein the composition comprises from about 0.5% to about 1.5% byweight of ionic silver.
 14. The antiseptic composition as claimed inclaim 1 wherein the oxidizing agent is sodium hypochlorite or chlorinedioxide.
 15. A method of making an antiseptic composition as claimed inclaim 4 comprising the steps of adding EDTA as a tetrasodium salt inaqueous solution and then adjusting the pH of the composition to pHabout 4 to about
 8. 16. A method of treating chronic wounds or burnscomprising applying to a chronic wound or a burn an antimicrobial agentand an agent which disrupts biofilms.
 17. The method as claimed in claim16 wherein the agent which disrupts biofilms is EDTA.
 18. The method asclaimed in claim 16 wherein the antimicrobial agent is selected from thegroup consisting of iodine, ionic silver or an oxidizing agent.
 19. Themethod as claimed in claim 18 wherein the oxidizing agent is sodiumhypochlorite or chlorine dioxide.